LOCUS       sequence-433923-        5790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DV4 stable E dimer with fusion loop mutation and increased 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5790)
  AUTHORS   Phan TTN, Hvasta MG, Kudlacek ST, Thiono DJ, Tripathy A, Nicely NI, 
            de Silva AM, Kuhlman B
  TITLE     A conserved set of mutations for stabilizing soluble envelope 
            protein dimers from dengue and Zika viruses to advance the 
            development of subunit vaccines.
  JOURNAL   J Biol Chem. 2022 Jul;298(7):102079. doi: 10.1016/j.jbc.2022.102079.
            Epub 2022 May 26.
  PUBMED    35643320
REFERENCE   2  (bases 1 to 5790)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem."; date: "2022-07"; volume: "298(7):102079. doi"; pages: " 
            10.1016/j.jbc.2022.102079. Epub 2022 May 26"
COMMENT     Sequence Label: pD4sE_SC12
FEATURES             Location/Qualifiers
     source          1..5790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             complement(1934..1945)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3020..3043
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3145..3164)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3210..3265
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3264..3283)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3626..3642)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3626..3642)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3639..3661)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3650..3666
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3674..3680,3681..3698,3699..3704))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3719..3740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3869..3886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4040..4628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4120..4139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4799..5590,5591..5659))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5422..5441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5660..5764)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrgggggg gggggggggg gggggggggg
      541 gggsggggsg rggsggrgrg gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gkgggggtgc cgggcggggc
     1261 ggggccgcck cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
     1741 tcaggccgag ttcggtaccg ccaccatgaa gtgggtaacc tttatttccc ttctttttct
     1801 ctttagctcg gcttattcca tgcggtgcgt cggcgtcggt aatagagatt ttgtagaggg
     1861 ggtatcagga ggtgcttggg tggatctggt gcttgagcat ggcaagtgcg taacggtgat
     1921 gatgcaaggt aaacctaccc tcgatttcga gttgactaag acgacagcaa aggaagtagc
     1981 gcttctcagg acctattgta tagaagcgtc aataagcaac ataaccacag cgacgcggtg
     2041 cccaactcag ggtgaaccct acttgaaaga ggagcaggac caacaatata tctgtagacg
     2101 agacgtcgta gatagggggt ggggaaatgg ctgtgacctg tttggcaagg gcggagtcgt
     2161 cacatgcgcc aaattctcat gctctggtaa gatcaccggt aatctggtgc aaatagaaaa
     2221 tcttgaatat acagtcgtgg tgacagttca caatggggac actcacgccg tcggcaacga
     2281 tactagtaac catggcgtta cagctatgat aactccccga tctccatcag tggaagtgaa
     2341 gttgccagat tatggagagc ttaccctcga ttgtgaacca agatccggta ttgacttcaa
     2401 tgaaatgata ttgatgaaaa tgaaaaagaa aacgtggctg gtgcacaagc agtggttctt
     2461 ggatctcccc ttgccctgga ctgcaggcgc ggacacgtcc gaggttcatt ggaactataa
     2521 agaaagaatg gttaccttca aagtcccgca cgcaaagaga caagacgtca ctgtcttggg
     2581 tagtcaggaa ggatggatgc accgggcact ggctggtgct actgaagttg actctggaga
     2641 tggaaaccat atgtggcccg ggcatctgaa gtgcaaggtt cgcatggaga agttgagaat
     2701 caagggtatg tcatatacga tgtgtagtgg aaaatttagc atagataaag agatggctga
     2761 aacacaacat ggaacgacag tcgttaaggt taagtatgag ggagctgggg ctccctgcaa
     2821 ggttcccata gaaatccggg acgttaataa agagaaagtc gtcgggcgca tcattagttc
     2881 cactccattg gctgagaaca caaattctgt gacaaacatt gagctggagc cgccgttcgg
     2941 ggactcatat atcgtcatag gagtgggtaa ttccgcgctg actttgcact ggttccgcaa
     3001 aggcagcagc ggcggcagcc atcaccacca tcatcaccat cattagtaat gactcgagcg
     3061 ataattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     3121 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     3181 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     3241 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     3301 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     3361 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3421 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3481 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     3541 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     3601 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3661 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3721 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3781 tgtcgtgcca gcggatcgat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3841 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3901 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3961 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4021 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4081 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4141 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4201 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4261 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4321 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4381 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4441 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4501 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4561 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4621 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4681 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4741 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4801 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4861 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4921 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4981 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5041 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5101 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5161 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5221 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5281 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5341 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5401 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5461 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5521 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5581 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5641 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5701 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5761 cgcgcacatt tccccgaaaa gtgccacctg
//
